Cell Signaling Technology

Product Pathways - NF-kB Signaling

IRF-8 (D20D8) Rabbit mAb #5628

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP ChIP H M (R) (Mk) (X) (B) Endogenous 50 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  X=Xenopus  B=Bovine
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

IRF-8 (D20D8) Rabbit mAb detect endogenous levels of total IRF-8 protein. An unknown background band is detected at 80 kDa in some cell lines.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly65 of human IRF-8 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from Raji and Jurkat cells, untreated or treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml; overnight) using IRF-8 (D20D8) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from THP-1, RL7, and A20 cell lines using IRF-8 (D20D8) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected or transfected with human and mouse IRF-8 constructs, using IRF-8 (D20D8) Rabbit mAb.


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 THP-1 and either 20 μl of IRF-8 (D20D8) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human GPR18 promoter primers, SimpleChIP® Human MS4A7 Promoter Primers #9013, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Background

Interferon regulatory factors (IRFs) comprise a family of transcription factors that function within the Jak/Stat pathway to regulate interferon (IFN) and IFN-inducible gene expression in response to viral infection (1). IRFs play an important role in pathogen defense, autoimmunity, lymphocyte development, cell growth, and susceptibility to transformation. The IRF family includes nine members: IRF-1, IRF-2, ISGF3γ/p48, IRF-3, IRF-4 (Pip/LSIRF/ICSAT), IRF-5, IRF-6, IRF-7, and IRF-8/ICSBP. All IRF proteins share homology in their amino-terminal DNA-binding domains. IRF family members regulate transcription through interactions with proteins that share similar DNA-binding motifs, such as IFN-stimulated response elements (ISRE), IFN consensus sequences (ICS), and IFN regulatory elements (IRF-E) (2).

IRF-8/ICSCP is expressed predominately in hematopoietic cells and is further increased upon treatment with interferon (3,4). IRF-8 can function as a transcription repressor of ICS-containing promoters (4). Expression of IRF-8 can lead to the down-regulation of the anti-apoptotic protein Bcl-2 (5). Originally described as being induced by IFN-γ, IRF-8 expression is also elevated by IRF-α as well as IL-12 in NK and T cells (6). IRF-8 deficient mice have enhanced susceptibility to various pathogens and impaired production of interferons, as well as deregulated hematopoiesis that resembles chronic myelogenous leukemia (7,8). IRF-8 also regulates bone metabolism by suppressing osteoclast formation (9).

  1. Taniguchi, T. et al. (2001) Annu Rev Immunol 19, 623-55.
  2. Honda, K. and Taniguchi, T. (2006) Nat Rev Immunol 6, 644-58.
  3. Driggers, P.H. et al. (1990) Proc Natl Acad Sci U S A 87, 3743-7.
  4. Weisz, A. et al. (1992) J Biol Chem 267, 25589-96.
  5. Burchert, A. et al. (2004) Blood 103, 3480-9.
  6. Lehtonen, A. et al. (2003) Cytokine 24, 81-90.
  7. Holtschke, T. et al. (1996) Cell 87, 307-17.
  8. Fehr, T. et al. (1997) J Exp Med 185, 921-31.
  9. Zhao, B. et al. (2009) Nat Med 15, 1066-71.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

Products